KR20060062734A - Oral micro-emulsion composition comprising tacrolimus - Google Patents

Abstract

The present invention relates to a stable oral microemulsion composition of poorly soluble tacrolimus with increased bioavailability, wherein the composition of the present invention comprising tacrolimus active ingredient, cosurfactant, surfactant, oil and organic acid Easily spontaneously emulsified in the excellent dissolution rate can greatly improve the bioavailability of tacrolimus upon oral administration.

Description

Oral microemulsion composition of tacrolimus {ORAL MICRO-EMULSION COMPOSITION COMPRISING TACROLIMUS}             

1 is a result of measuring the particle size distribution during emulsification of the preparation according to the present invention,

Figure 2 is a graph comparing the absorption rate (change in blood concentration with time) upon oral administration of the preparation according to the invention and the control formulation (Prograf ^R , Fujisawa Island).

The present invention relates to an oral microemulsion composition of poorly soluble Tacrolimus, with increased bioavailability.

Tacrolimus is a macrolide antibiotic that has been found in fermentation broth of Streptomyces tsukubaensis and has an immunosuppressive effect. Although structurally different from cyclosporin A (CsA), the most widely used immunosuppressive agent, it is known that its mechanism of action is similar. Tacrolimus (FK506) binds to the intracellular tacrolimus binding protein FKBP (FK506 binding protein) 12 to form a FK506-FKBP complex. The complex binds to a serine / threonine dephosphatase called calcineurin, which inhibits the function of the enzyme, which in turn inhibits T cell activation. Tacrolimus is used as a primary or palliative therapy for tissue rejection reactions during kidney or liver transplantation. It is evaluated to have higher efficacy than A.

However, tacrolimus is very poorly soluble in water (approximately 1.58 μM in water at 25 ° C.), has very low elution, and thus has very low bioavailability when taken orally, and the deviation range of utilization is also very large, 5 to 65%.

Thus, as a method for increasing the solubility of the tacrolimus component, Korean Patent Publication No. 199595210 discloses a method for preparing a solid dispersion using a water-soluble polymer such as hydroxypropyl methylcellulose. However, this method is known to have many problems such as low reproducibility and complicated manufacturing process, such as evaporating and removing organic solvents after homogeneous suspension of tacrolimus, water-soluble polymer, lactose and sodium croscarmellose.

In addition, U. S. Patent No. 5,260, 301 discloses a solution formulation using ethanol as a solvent to increase the solubility and stability of tacrolimus, but when formulated into an extended capsule or the like, the ethanol volatilizes over time or volatilizes through a gelatinous coating even after preparation. This results in a deviation from the desired initial composition ratio, and as a result, the drug is separated from the precipitation or the composition ratio of forming a microemulsion has a disadvantage.

Accordingly, the present inventors have completed the present invention by discovering that oral microemulsion compositions containing tacrolimus exhibit thermodynamic stability, solubility and high bioavailability than conventional formulations.

Accordingly, it is an object of the present invention to provide a stable oral microemulsion composition containing tacrolimus with increased bioavailability.

In accordance with the above object, the present invention provides a tacrolimus-containing oral microemulsion composition comprising tacrolimus as an active ingredient and a cosurfactant, a surfactant, an oil and an organic acid.

Hereinafter, the present invention will be described in detail.

Since the composition of the present invention is a microemulsion state in which the poorly soluble tacrolimus component is completely dissolved, it is excellent in stability such as not forming crystals even in long-term storage, and is easily emulsified in biological fluids, and shows excellent dissolution rate by oral administration. The active ingredient can be usefully absorbed in vivo.

Hereinafter, it demonstrates according to the characteristic and the kind of each component which contain the composition of this invention.

(1) Active Ingredient

In the present invention, tacrolimus is used as the active ingredient.

(2) co-surfactant

Cosurfactants necessary for the emulsification of solubilizers and preparations used for the purpose of dissolving poorly water-soluble tacrolimus are non-toxic transcutol (dicutane glycol diethyl glycol monoethyl ether), polyethylene glycol (molecular weight 200 to 600), tria Cetine and mixtures thereof can be used.

In acute oral toxicity studies in mice, transcutol had a LD ₅₀ of 7.95 ml (weight 0.989) / kg (Gattefosse product profile) and polyethylene glycol had a LD ₅₀ of 28.9 g / kg (Handbook of pharmaceutical excipients, p570-571, 3rd Ed., American pharmaceutical association, Washington, DC) .A low-toxic, stable soft-capsule formulation with lower reactivity with gelatinous coatings than that produced with hydrophilic cosurfactants such as propylene glycol, ethanol, or propylene glycol monoacetate. Can be prepared. Triacetin is used as a polymer coating for capsules, tablets, and granules. It has been reported to be used as a solvent for food, and to be used as a nitrogen source after metabolism in parenteral fluid preparations, and to be used for solubilizing Taxol, a poorly soluble anticancer agent.

(3) surfactant

As the surfactant of the present invention, any pharmaceutically acceptable surfactant which can form a stable microemulsion by stably emulsifying hydrophilic components such as oil and cosurfactant in water can be used. Same as

① reaction products of natural or hydrogenated vegetable oils with ethylene glycol, ie polyoxyethylene glycolated natural or hydrogenated vegetable oils, such as polyoxyethylene glycolated natural or hydrogenated castor oil (trade name: Cremophor, BASF), HCO, Nikkol),

② polyoxyethylene-sorbitan-fatty acid esters, for example, mono, trilauryl, palmityl, stearyl, oleyl ester (product name: Tween, ICI),

③ polyoxyethylene fatty acid esters, for example polyoxyethylene stearic acid ester (product name: Myrj, ICI),

④ polyoxyethylene-polyoxypropylene block copolymer (trade name: Poloxamer, Pluronic, Lutrol; BASF),

⑤ mono-, di- or mono / di-glycerides, for example capryl / capric acid mono- or di-glycerides (trade name: Imwitor, Huls),

⑥ sorbitan fatty acid esters, for example sorbitan monolauryl, sorbitan monopalmityl or sorbitan monostearyl (trade name: Span),

⑦ trans-esterification products of natural vegetable oil triglycerides and polyalkylene polyols (trade name: Labrafil, Gattefosse).

These may each be used alone or as a mixture of two or more thereof, preferably cremophore which is a polyoxyethylene product of hydrogenated vegetable oil or tween which is polyoxyethylene-sorbitan-fatty acid ester, vegetable oil triglyceride and polyalkyl. Labrafil, which is the product of trans-esterification of ene polyols, can be used.

(4) oil components

In the present invention, the oil component may be any pharmaceutically acceptable oil that is well mixed with the surfactant and stably emulsified in water to form a stable microemulsion, and representative examples thereof are as follows.

① fatty acid triglycerides, preferably intermediate fatty acid triglycerides, for example fractionated coconut oil (trade name: Miglyol, Huls, Captex, Abitec),

② mono-, di- or mono / di-glycerides, preferably mono- or di-glycerides of oleic acid,

(3) ester compounds of fatty acids and monovalent alkanols, in particular ester compounds of fatty acids having 8 to 20 carbon atoms and monovalent alkanols having 2 to 3 carbon atoms, for example isopropyl myristate, isopropyl palmitate, ethyl linoleate, ethyl Orleans,

Propylene glycol mono- or di-fatty acid esters such as propylene glycol dicaprylate, propylene glycol monocaprylate, propylene glycol dilaurate, propylene glycol isostearate, propylene glycol monolaurate, propylene glycol ricinoli Eight,

⑤ hydrocarbons, for example squalene, squalane,

(6) tocopherols such as tocopherol, tocopherol acetate, tocopherol succinate, polyethylene glycol-1000-tocopherol succinate (TPGS), and the like.

These may each be used alone or as a mixture of two or more thereof, preferably of these, ethyl linoleate, which is an ester compound of fatty acid and monovalent alkanol, migliol, captex, mono-, di- or fatty acid triglyceride. Mono / di-glycerides, tocopherols can be used.

(5) organic acids

In the present invention, a pharmaceutically acceptable organic acid is added to the composition to solve the stability problem for tacrolimus in the preparation of a microemulsion for oral administration. Examples of representative organic acids include erythorbic acid, citric acid, tartaric acid, ascorbic acid. , Lactic acid, malic acid, succinic acid, acetic acid, trichloroacetic acid, trifluoroacetic acid, dimethyltriaminepentaacetic acid, pyruvic acid, malonic acid, myristic acid, picric acid, methanesulfonic acid, ethanesulfonic acid, p-aminobenzoic acid, benzenesulfuric acid Phonic acid, benzoic acid, edetic acid, sorbic acid, adipic acid, gluconic acid, aminocaproic acid, glycidic acid, isostearic acid, dodecylbenzenesulfonic acid, fumaric acid, maleic acid, oxalic acid, butyric acid, palmitic acid, sulfonic acid, sulfinic acid, Formic acid, propionic acid, tannic acid, pantothenic acid, aspartic acid, aminoacetic acid, DL-α-aminopropionic acid, and the like.

These may each be used alone or as a mixture of two or more, preferably erythorbic acid or citric acid may be used.

The composition of the present invention comprises tacrolimus, co-surfactant, surfactant, oil, and organic acids, which are the aforementioned constituents, from 1: 5 to 200: 5 to 400: 1 to 100: 0.01 to 50, preferably from 1 to 10 to 150. 10 to 300: 2 to 80: 0.1 to 20 in weight ratio.

The oral microemulsion composition of the present invention can be prepared by dissolving the tacrolimus component homogeneously in a cosurfactant as a solvent, and then adding a surfactant, oil and organic acids to the solution to dissolve it, and the average diameter when contacted with an aqueous solution Fine emulsified particles of 1 μm or less are formed. The composition thus prepared may be formulated by filling in hard capsules or soft capsules according to a conventional pharmaceutical manufacturing process.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following examples are only for illustrating the present invention, and the scope of the present invention is not limited thereto.

Example 1

After tacrolimus was dissolved in transcutol, the remaining components were added and dissolved in turn to obtain a microemulsion preconcentrate. This preconcentrate was formulated by filling into soft capsules according to the method for producing soft capsules in the Korean Pharmaceutical Formulation.

Comparative Example 1

It was formulated in the same manner as in Example 1 using the ingredients shown in Table 2 in the corresponding amounts.

Comparative Example 2

It was formulated in the same manner as in Example 1 using the ingredients shown in Table 3 in the corresponding amounts.

Example 2

It was formulated in the same manner as in Example 1 using the ingredients shown in Table 4 below in the corresponding amounts.

Example 3

It was formulated in the same manner as in Example 1 using the ingredients shown in Table 5 in the corresponding amounts.

Example 4

It was formulated in the same manner as in Example 1 using the ingredients shown in Table 6 below in the corresponding amounts.

Example 5

It was formulated in the same manner as in Example 1 using the ingredients shown in Table 7 in the corresponding amounts.

Example 6

It was formulated in the same manner as in Example 1 using the ingredients shown in Table 8 in the corresponding amounts.

Example 7

It was formulated in the same manner as in Example 1 using the ingredients shown in Table 9 in the corresponding amounts.

Example 8

It was formulated in the same manner as in Example 1 using the ingredients shown in Table 10 in the corresponding amounts.

Example 9

It was formulated in the same manner as in Example 1 using the ingredients shown in Table 11 in the corresponding amounts.

Example 10

It was formulated in the same manner as in Example 1 using the ingredients shown in Table 12 in the corresponding amounts.

Example 11

It was formulated in the same manner as in Example 1 using the ingredients shown in Table 13 in the corresponding amounts.

Example 12

It was formulated in the same manner as in Example 1 using the components shown in Table 14 below in the corresponding amounts.

Example 13

It was formulated in the same manner as in Example 1 using the ingredients shown in Table 15 below in the corresponding amounts.

Example 14

It was formulated in the same manner as in Example 1 using the ingredients shown in Table 16 in the corresponding amounts.

Example 15

It was formulated in the same manner as in Example 1 using the ingredients shown in Table 17 below in the corresponding amounts.

Example 16

It was formulated in the same manner as in Example 1 using the ingredients shown in Table 18 in the corresponding amounts.

Example 17

It was formulated in the same manner as in Example 1 using the ingredients shown in Table 19 below in the corresponding amounts.

Example 18

It was formulated in the same manner as in Example 1 using the ingredients shown in Table 20 in the corresponding amounts.

Example 19

It was formulated in the same manner as in Example 1 using the ingredients shown in Table 21 below in the corresponding amounts.

Example 20

It was formulated in the same manner as in Example 1 using the ingredients shown in Table 22 below in the corresponding amounts.

Test Example 1 Analysis of Particle Size Distribution of Emulsified Particles

0.1 g of the formulation prepared in Example 1 was diluted in 10 ml of purified water, and then the particle size distribution was measured using a particle analyzer (Shimadzu, SALD-2001 model, Japan), and the results are shown in FIG. 1.

From Figure 1, it can be seen that the preparation prepared according to the present invention easily forms a microemulsion by forming fine emulsion particles having an average particle diameter of less than about 1 ㎛ when contacted with an aqueous solution.

Test Example 2: Precipitation Formation Test

In order to confirm the formation of precipitates upon contact with purified water, artificial gastric juice and artificial intestinal fluid of the formulation prepared in Example 1, 0.1 g of the formulation was diluted in 10 ml of the solution, and then visually observed whether precipitates were formed (+: precipitation generation, -: No precipitation is produced), the results are shown in Table 23 below.

As shown in the results of Table 23, the formulation prepared according to the present invention does not cause precipitation upon contact with the bioaqueous solution.

Test Example 3: Stability Test

As a test formulation, the formulations prepared in Comparative Examples 1, 2 and Example 6 and 1 mg of commercially available hard capsule formulation Prograb (Prograf ^R , Fujisawa Island) as a control formulation were stored at 60 ° C. and then over time. The content of was analyzed by HPLC to show the stability of the formulation in Table 24.

Column: Inertsil ODS2 (4.6 X 250 mm)

Mobile phase: Acetonitrile: Distilled water: Phosphoric acid (60: 40: 1)

Injection volume: 10 μl

Oven: 50 ℃

UV detector: 210 nm

As shown in the results of Table 24, in the case of Example 1 to which the organic acid was added, it is more stable in the harsh conditions than in the case of Comparative Example 1 to which the organic acid was not added. Solid formulations, such as the control formulation, can secure some stability, but when formulated into a microemulsion by solution, the thermodynamic activity is high, which may cause stability problems. Therefore, by using organic acids such as erythorbic acid or citric acid, It can be seen that as in Example 1 to ensure the stability of the formulation.

Test Example 4 Comparative Absorption Test

The bioavailability comparison test of rats when orally administered using the preparation prepared in Example 1 as a test formulation and 1 mg of prograb which is a commercially available hard capsule formulation as a control formulation was carried out as follows.

As the experimental animals, three Sprague-Dawley male rats (250 g in weight and 14-15 weeks old) were used for each sample. The animals were fed with water. Rats were fasted for more than 48 hours and used for the test. During fasting, water was allowed to drink freely.

Rats were dosed with a test or control formulation with a water dose of 10 mg of tacrolimus per kilogram of rat weight with water using an oral administration device, 0.5, 1, 1.5, 2, 3, 4 before and after administration. Blood was collected at, 5, 7, and 24 hours later.

400 μl of a methanol solution containing zinc sulfate (0.2 M ZnSO ₄ : MeOH = 2: 8) was added to 200 μl of the collected blood, followed by mixing and shaking. The solution was centrifuged at 12,000 rpm for 30 seconds, the supernatant was collected, filtered through a 0.22 μm filter, and analyzed using LC-MS. The results are shown in Table 25 and FIG. 2.

Column: Waters MS C18 (2.1 × 150 mm, equipped with guard column)

Mobile phase: 65% methanol, 95% methanol

Injection volume: 30 μl

Flow rate: 0.3 ml / min

Detection: SIR Mode m / z: 826.7 (Na adduct)

From Table 25 and Figure 2, it can be seen that the formulation prepared according to the present invention shows an increased bioavailability of about 2 times or more compared to the control formulation prograb.

The composition of the present invention is a microemulsion state in which poorly soluble tacrolimus component is completely dissolved in a solvent having low toxicity, and does not form crystals even during long-term storage. Oral administration can greatly improve the bioavailability of tacrolimus.

Claims (6)

A tacrolimus-containing oral microemulsion composition comprising tacrolimus, a cosurfactant, a surfactant, an oil and organic acids as active ingredients. The method of claim 1, Tacrolimus: Co-surfactant: Surfactant: Oil: Organic acids are contained in the weight ratio of 1: 5 to 200: 5 to 400: 1 to 100: 0.01 to 50. The method of claim 1, The cosurfactant is selected from the group consisting of transcutol, polyethylene glycol, triacetin and mixtures thereof. The method of claim 1, The surfactant is selected from the group consisting of polyoxyethylene products of hydrogenated vegetable oils, polyoxyethylene-sorbitan-fatty acid esters, trans-esterification reaction products of natural vegetable oil triglycerides and polyalkylene polyols, and mixtures thereof The composition characterized in that. The method of claim 1, The oil is selected from the group consisting of ester compounds of fatty acids and monohydric alkanols, fatty acid tri-, mono-, di- or mono / di-glycerides, tocopherols and mixtures thereof. The method of claim 1, Organic acids include erythorbic acid, citric acid, tartaric acid, ascorbic acid, lactic acid, malic acid, succinic acid, acetic acid, trichloroacetic acid, trifluoroacetic acid, dimethyltriaminepentaacetic acid, pyruvic acid, malonic acid, myristic acid, picric acid, methane Sulfonic acid, ethanesulfonic acid, p-aminobenzoic acid, benzenesulfonic acid, benzoic acid, edetic acid, sorbic acid, adipic acid, gluconic acid, aminocaproic acid, glycidic acid, isostearic acid, dodecylbenzenesulfonic acid, fumaric acid, maleic acid , Oxalic acid, butyric acid, palmitic acid, sulfonic acid, sulfinic acid, formic acid, propionic acid, tannic acid, pantothenic acid, aspartic acid, aminoacetic acid, DL-α-aminopropionic acid, and the like. Composition.

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